Biomedical subjects
R Palmiter
Publications and source records attributed to R Palmiter.
Norepinephrine is required for leptin effects on gene expression in brown and white adipose tissue.
Exogenous leptin enhances energy utilization in ob/ob mice by binding its hypothalamic receptor and selectively increasing peripheral fat oxidation. Leptin also increases uncoupling protein 1 (UCP1) expression in brown adipose tissue (BAT), but the neurotransmitter that mediates this effect has not been established. The present experiments sought to determine whether leptin regulates UCP1 expression in BAT and its own expression in white adipose tissue (WAT) through the long or short forms of leptin receptor and modulation of norepinephrine release. Mice lacking dopamine beta-hydroxylase (Dbh-/-), the enzyme responsible for synthesizing norepinephrine and epinephrine from dopamine, were treated with leptin (20 microg/g body weight/day) for 3 days before they were euthanized. UCP1 messenger RNA (mRNA) and protein expression were 5-fold higher in BAT from control (Dbh+/-) compared with Dbh-/- mice. Leptin produced a 4-fold increase in UCP1 mRNA levels in Dbh+/- mice but had no effect on UCP1 expression in Dbh-/-. The beta3-adrenergic agonist, CL-316,243 increased UCP1 expression and established that BAT from both groups of mice was capable of responding to beta-adrenergic stimulation. Similarly, exogenous leptin reduced leptin mRNA in WAT from Dbh+/- but not Dbh-/- mice. In separate experiments, leptin produced comparable reductions in food intake in both Dbh+/- and Dbh-/- mice, illustrating that norepinephrine is not required for leptin's effect on food intake. Lastly, db/db mice lacking the long form of the leptin receptor failed to increase UCP1 mRNA in response to exogenous leptin but increased UCP1 mRNA in response to CL-316,243. These studies establish that norepinephrine is required for leptin to regulate its own expression in WAT and UCP1 expression in BAT and indicate that these effects are likely mediated through the centrally expressed long form of the leptin receptor.
The effect of thymus environment on T cell development and tolerance.
During development in the thymus, T cells are deleted if their receptors are able to recognize self major histocompatibility complex (MHC) proteins. We show that such clonal deletion can occur because of interaction between receptors on T cells and MHC expressed on bone marrow-derived cells. In addition, development in the thymus picks out T cells to mature if their receptors will be restricted for antigen recognition in association with self MHC alleles expressed on thymus epithelial cells. This process is usually thought to involve positive selection of T cells bearing receptors with high and low affinity for MHC on thymus epithelium, and subsequent deletion of high affinity cells by interaction with bone marrow-derived cells. Our data do not fit such a model, but rather suggest that MHC molecules on thymus epithelium and bone marrow-derived cells may not be seen identically by T cell receptors.
Progressive glomerulosclerosis develops in transgenic mice chronically expressing growth hormone and growth hormone releasing factor but not in those expressing insulinlike growth factor-1.
An increase in glomerular size occurs in normal maturation after subtotal renal ablation and disease states such as diabetes mellitus. The role that growth hormone (GH), growth hormone releasing factor (GHRF), and insulinlike growth factor-1 (IGF-1) play in these processes has been investigated using transgenic mice chronically expressing these hormones. The glomeruli were enlarged in all 3 strains of mice. Mesangial proliferation followed by progressive glomerulosclerosis was observed in the GH and GHRF animals only. In the IGF-1 mice the large glomeruli remained morphologically normal except for the enlargement. These data suggest that the glomerulosclerosis was due, in part, to disordered mesangial cell growth in response to circulating GH. The mesangial lesions in mice with chronically high plasma GH levels mimicked those in human diabetes mellitus. These models provide a means to study the hormonal regulation of glomerular growth and the role that specific hormones might play in the pathogenesis of glomerulosclerosis.
Secretory rhythm of growth hormone regulates sexual differentiation of mouse liver.
The secretory pattern of growth hormone (GH) differs between the sexes; in males it is more pulsatile than in females. Experiments were performed to test the hypothesis that differences in the secretory rhythm of GH are responsible for sex-dependent liver functions of mice. Continuous delivery of GH was achieved either by introducing metallothionein-GH fusion genes into the germ line or by implanting minipumps. Pulsatile delivery of GH was mimicked by injection. The effects of these treatments on production of hepatic prolactin/GH receptors, albumin, and major urinary protein (MUP) were monitored. The results suggest that induction of MUP mRNA requires pulsatile occupancy of GH receptors, which is achieved naturally in males or by injection of GH, whereas chronic occupancy of GH receptors is inhibitory. In contrast, induction of prolactin/GH receptors requires chronic stimulation of GH receptors, which is approximated in normal female mice or results from increased GH levels in mice with foreign genes or undergoing infusions from minipumps.
The ovalbumin gene family: hormonal control of X and Y gene transcription and mRNA accumulation.
The ovalbumin gene family is composed of three genes, X, Y and ovalbumin, which are expressed in laying hen oviduct. We have analyzed the in vivo transcription products of X and Y genes and the effect of steroid hormones on their synthesis and accumulation. As in the case of ovalbumin, the complete gene transcripts and processing intermediates are present in the poly(A)+ RNA fraction. The mature RNAs are found in polysomes and are translated into proteins. The expression of X and Y genes is controlled by steroid hormones: X and Y RNAs are not detectable in oviducts from chicks withdrawn from estrogen stimulation, whereas in chicks stimulated with estrogen for 7 days, X RNA represents 0.3% and Y RNA 0.8% of ovalbumin mRNA. In laying hen, however, the levels of X and Y RNAs are about 2% of ovalbumin mRNA. After stimulation with other steroid hormones, alone or in combination, the level of X and Y RNA does not achieve that detected in laying hen. Progesterone has a much weaker effect on X RNA accumulation than on that of Y and ovalbumin mRNAs. Studies with isolated nuclei show that X and Y gene expression is regulated by hormones at the level of transcription. However, the differences observed between the transcription rates and the accumulation of X and Y mRNAs suggest that the expression of X and Y genes could also be controlled at the levels of RNA processing and/or mRNA stability.
Cloning of an almost full-length chicken conalbumin double-stranded cDNA.
Chicken conalbumin double-stranded cDNA (con-dscDNA) was synthesized from a laying hen oviduct mRNA preparation enriched for conalbumin mRNA (con-mRNA). The dscDNA was inserted by blunt-end ligation into the Sal I site of plasmid pBR322 which had been repaired with DNA polymerase I to create Taq I sites on each side of the inserted fragment. After bacterial transformation, one hybrid recombinant, pBR322-con1, which contains the largest inserted dscDNA (about 2350 bp) was shown to hybridize specifically to the RNA which is translated into conalbumin. Electron microscopic examination of hybrid molecules between con-mRNA and pBR322-con1 DNA indicate that the inserted con-dscDNA is an almost full-length double-stranded transcript of conalbumin mRNA.